Biotechnological process for hydrocarbon recovery in low permeability porous media

ABSTRACT

The present invention refers to a biotechnological process that enhances oil recovery of a 14 to 25 API Gravity oil contained in carbonate-containing and/or clayey sandstone porous rock systems with low permeability (7 to 100 mD), thus focused to petroleum wells associated to zones with low recovery factor. The process utilizes the indigenous extremophile microorganisms activity from the oil reservoir and an IMP culture, as well as its metabolites (gases, acids, solvents and surfactants), which improve oil mobility and are able to develop at 60 to 95° C. temperatures, 7 to 154.6 Kg/cm 2  (100 to 2,200 psi) pressures and NaCl content from 5,000 to 45,000 ppm, in anaerobic conditions. The biotechnological process of the present invention includes: indigenous microorganisms sampling, collecting and characterization from the reservoir; formulation of culture media; selection, enrichment, activation and preservation of such microorganisms, as well as biostimulation (indigenous microorganisms) and bioaugmentation (IMP culture) to increase the metabolite production, useful for oil recovery in a porous media impregnated with oil in carbonate-containing and/or clayey sandstone porous systems, with one or several cycles and confinement periods from 5 to 10 days, to increase oil recovery. Experimental tests show that the biotechnological process of the present invention supplies an oil recovery increase up to 30%, additional to that obtained in secondary recovery processes.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit under 35 U.S.C. §119 of Mexican Patent Application No. MX/a/2009/014146, filed Dec. 21, 2009, which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to a biotechnological process for increasing hydrocarbon recovery with an API Gravity from 14 to 25, contained in carbonate-containing and/or clayey sandstone porous systems, with low permeability. The invention is also directed to increasing recovery from petroleum wells and associated zones with low hydrocarbon recovery factor using the activity of indigenous extremophile microorganisms from the oil reservoir, as well as its metabolites to help oil mobility, that are able to develop at temperatures ranging from 60 to 95° C., pressures from 7 to 154.6 Kg/cm² (100 to 2,200 psi) and NaCl content from 5,000 to 45,000 ppm, in anaerobic conditions.

BACKGROUND

Nowadays 85% of the world crude oil production is by means of primary and secondary recovery, with an approximate 35% recovery of the petroleum existing in the reservoir (Babadagli T., 2007: Development of mature oil fields—A review. J. Petroleum Science Engineering 57: 221-246). Because the recovery rate is considered low, other enhanced recovery methods and systems of petroleum have been implemented, EOR (Enhanced Oil Recovery).

Oil Recovery can be classified into three processes:

-   1. Primary recovery: Oil is produced from the geological formation     towards the well by only using the energy from the oil or gas     reservoir and/or forcing it through pumping. -   2. Secondary recovery: consists on an increase of pressure in the     reservoir by means of an energy from an external source; fluid     injection in the rock of the reservoir is a common process to carry     higher volumes of petroleum to the surface; over 25% of petroleum is     recovered this way. -   3. Tertiary or EOR: Oil recovery from a reservoir, above the one     recovered by means of primary or secondary recovery; normally     implying a method such as reservoir heating or using chemicals. The     kind of procedures for tertiary recovery include injecting to the     reservoirs, miscible solvents, hydrocarbonated gases or carbonic     gas, as well as soda water, tensoactives or hydrosoluble polymers,     including any type of process used to supply the reservoir of an     additional source of energy to enhance petroleum recovery compared     to that produced during primary and secondary recovery processes.

Reservoir production starts with the natural flow of crude due to the energy in the reservoirs, being later completed by means of water and gas injection, like in the secondary recovery processes. The tertiary or EOR methods are not widely used because of the high costs and complexity in the handling of equipment. All the processes are used to maximize the economic value on a long term basis in hydrocarbon reserves application is scarce as resources are needed for its commercial exploitation.

The scarce recovery of petroleum in the reservoirs is due to several factors: low permeability, high viscosity of oil, high interfacial tension within water and petroleum, and high capillarity forces, resulting into low mobility (Chisholm et al., 1990: Microbial enhanced oil recovery: interfacial tension and gas-induced relative permeability effects. SPE: 20481, 8).

Because secondary recovery processes have economic and/or practical limitations in the application, the development of innovating technologies is required to implement exploitation processes (Planckaert M., 2005: Oil reservoirs and oil production. In Petroleum Microbiology. Ollivier B. y Magot M. (Ed.), ASM Press, Washington D.C. 3-19).

There are tertiary or EOR processes used to increase the productive life of wells; including water or other liquid injection to force the oil towards the production wells. The properties of the oil can be modified by adding surfactants that can in turn modify the wettability of the rock with oil (Mukherjee et al., 2006: Towards commercial production of microbial surfactants. Trends Biotechnol. 24(11):509-15; Singh et al., 2007: Surfactants in microbiology and biotechnology: Part 2. Application and aspects. Biotechnology Advances 25:99-121; Youssef et al., 2007: In situ biosurfactant production by Bacillus strains injected into a limestone petroleum reservoir. Appl. Environ. Microbiol. 73(4):1239-47).

Among the tertiary or EOR technologies are hydrocarbon recovery by means of microbial processes intended to solve this problem with the application of microorganisms and/or its metabolic products. This technology is environmental, technical and economically efficient, being useful to increase the production of oil in wells and reservoirs (Maure et al., 1999: Microbial enhanced oil recovery pilot test in Piedras Coloradas field, Argentina. SPE 53715; Bryant R and Lockhart T. P., 2002: Reservoir engineering analysis of microbial enhanced oil recovery. SPE Reservoir Evaluation & Engineering, 365-374; Nazina et al., 2008: Regulation of geochemical activity of microorganisms in a petroleum reservoir by injection of H₂O₂ or water-air mixture. Microbiology 77(3): 324-333).

Extremophile microorganisms can live in environments considered unsuitable for most organisms, such as high temperature, salinity and pressure, characteristics of petroleum reservoirs. Therefore the indigenous microorganisms from the reservoirs are the best ones to survive under these conditions. (Monastersky R., 1997: Deep dweller microbes thrive far below ground. Science 151(13):192-193).

International Patent Application No. WO 2009/009382 A3 “Process for enhanced oil recovery using a microbial consortium” Published on Jan. 15, 2009, Soni et al. proposes a method including the selection of the well wherein the process of hydrocarbon recovery uses a microbial consortium. The characteristics considered for the application of such a process are: well depth<3,048 m (10,000 ft), temperature<90° C., pressure<300 Kg/cm²; pH from 4 to 9, preferably from 6 to 8, total salinity of formation water<12% (<10% NaCl). Microbiological prospecting and the selection of microorganisms is carried out by using 4 different culture media, including nutrients, minerals and carbon source. This process comprises the injection of the media and microorganisms in the selected well, closing of the well from one to three weeks for the growth of the microbial consortium to enable the microorganisms to release the oil of the well, increasing the recovery of same.

U.S. Published Patent Application No. 20070092930 A1 entitled “Process for enhanced recovery of crude oil from oil wells using novel microbial consortium”, Published on Apr. 26, 2007 by Lal et al. discloses a process for hydrocarbon recovery, wherein a mixed culture of thermophiles, barophiles and acidogenic anaerobe bacteria, are developed in culture media containing nutrients, minerals and complex carbon source. The incubation and enrichment of the microorganisms is carried out at temperatures from 70 to 90° C. The microorganisms are obtained from samples collected under strict anaerobic conditions. During the enrichment of the samples the production of CO₂, CH₄ biosurfactants, fatty acids and alcohol, metabolites indicating the development of microorganisms of interest are monitored. Recovery examples were carried out in packed granular systems and Berea sandstone core, with a 148 mD permeability, 19 porosity and a 27.9% residual saturation, with an 8% oil recovery.

U.S. Published Patent Application No. 20070181300 A1 entitled “System and method for preparing near-surface heavy oil for extraction using microbial degradation”, Published on Aug. 9, 2007, by Bushe and Rollins discloses a heavy oil recovery system by means of bacteria and fungi, with the addition of nutrients. The application field of this patent is oil located near the reservoir surface, implying that the invention is used at temperatures and pressures relatively low or from the surface. The oil mobility mechanisms include the decrease of viscosity, metabolites production such as gases and acids and hydrocarbon biodegradation. The patent application does not disclose the oil densities interval or API Gravity for the applicability of the invention.

U.S. Pat. No. 5,163,510 entitled “Method of microbial enhanced oil recovery”, issued Nov. 17, 1992, to Sunde discloses a method to enhance oil recovery in rock by using exogenous aerobic microorganisms and nutrients (N and P) addition. Experiments describe the use of cores with 700 mD permeability and 45° C. systems incubation, obtaining a residual oil recovery of 9% after the secondary recovery. The characteristics of the oils to which the process can be applied is not specified. Mechanisms attributable to recovery are: CO₂ and biosurfactants production, as well as hydrocarbon degradation.

In many of the stated references, oil recovery using microorganisms has been carried out in high permeability systems such as Berea sandstone, with a 148 mD value or in systems with higher permeability (packed systems). On the other hand, such references do not disclose the permeability of the system used, or other important characteristics of the oil such as viscosity, API Gravity, pressure conditions, or referring that the microorganisms are resistant to high pressure conditions.

There is a continuing need for technologies focused on low permeability systems (lower than 100 mD), high pressure (up to 154.6 Kg/cm², 2,200 psi) and high temperatures (above 60° C.), conditions prevailing in many of the Mexican fields.

SUMMARY OF THE INVENTION

The prior technologies do not supply in the application field an alternative of solution to the oil recovery problem with an API gravity of up to 14 and found in porous media with permeability lower than 100 mD. The present invention is directed to a biotechnological process for enhancing oil recovery in an API Gravity of 14 to 25, contained in carbonate-containing rock and/or clayey sandstone porous systems with low permeability of up to 7 mD. The invention is particularly directed to petroleum wells associated to zones with low oil recovery factor through the stimulated activity of indigenous extremophile microorganisms from the oil well, in addition to its metabolites, which increase the mobility of oils.

One aspect of the present invention is to provide a biotechnological process to enhance hydrocarbon recovery located in carbonate-containing rock and/or clayey sandstone systems with 7 to 100 mD permeability.

An additional feature of the present invention is to provide a biotechnological process to enhance hydrocarbon recovery, using extremophile microorganisms: thermophile, barophile, halotolerant, acidotolerant and anaerobe, indigenous of the oil reservoirs; as well as its metabolites: surfactants, gases, acids and solvents, increasing in the mobility of oil and that can develop at temperatures ranging from 60 to 95° C. pressures from 7 to 154.6 Kg/cm² (100 a 2,200 psi) and NaCl content from 5,000 to 45,000 ppm, in anaerobic conditions. In one embodiment, the microorganisms are thermophilic, barophilic, acidogenic and anaerobic.

One feature of the process of the invention is to provide a process for increasing recovery of oil from porous rock or strata that can not be easily recovered by other methods. The process is particularly suitable for use with oil having 14 to 25° API. The process basically obtains a sample of the rock and fluids from the well where the fluids generally include oil and water. The rock sample is analyzed for its porosity and composition. The oil is analyzed for its API. The fluid sample is analyzed for microbial content and the well is analyzed for the temperature, pressure and salinity of the environment from which the microorganisms were found. A culture media is then obtained for culturing the microorganisms obtained from the well to obtain an indigenous microorganism culture. In one embodiment, the indigenous microorganisms are anaerobic that are capable of growing at temperatures of 60 to 95° C., salinity of 5,000 to 45,000 ppm NaCl, a pressure of 7 to 154.6 Kg/cm² (100 to 2,200 psi) and can grow in the presence of oil of 14 to 25° API such as that found in the well environment.

A culture media is then prepared containing one or more of the microorganisms selected from the group consisting of Thermoanaerobacter ethanolicus, Thermoanaerobacter uzonensis, Thermoanaerobacter inferii, Geothermobacterium sp, Methanobacterium subterraneum, Methanobacterium formicicum, Methanolinea tarda and/or Methanoculleus sp, and mixtures thereof. An inoculum is then prepared from a mixture of the indigenous microorganisms and the culture media which is then injected into the well at a pressure of 7 to 154.6 Kg/cm² and at a temperature of 60 to 95° C. After a period of time, the oil is recovered from the well. The indigenous microorganisms and the culture media are combined in a ratio to treat the oil well effectively to enhance recovery of the oil from the well.

Another feature of the invention is to provide a process of injecting a mixture of indigenous microorganisms and the culture media into a porous rock containing heavy oil of 7-25° API. The mixture can also contain a nutrient growth media for promoting the growth and culturing of the microorganisms. The nutrient growth media can be included in an amount effective to culture the microorganisms in amounts to treat the oil to enhance oil recovery from the well. The mixture is injected into the well for a time and at a temperature sufficient to enable the microorganisms to grow and reduce the viscosity of the oil whereby the oil can be recovered from the well.

Another feature of the invention is directed to a process that is able to decrease the content of the aromatic compounds in the oil and convert the aromatic compounds to lower molecular weight compounds such as aliphatic or lower molecular weight aromatic compounds. The process includes the step of injecting a mixture of cultured indigenous microorganisms obtained from the well and one or more microorganisms selected from the group consisting of Thermoanaerobacter ethanolicus, Thermoanaerobacter uzonensis, Thermoanaerobacter inferii, Geothermobacterium sp, Methanobacterium subterraneum, Methanobacterium formicicum, Methanolinea tarda and/or Methanoculleus sp, and mixtures thereof. The culture mixture can contain a nutrient media. The microorganisms produce metabolites that in conjunction with the pressure and temperature in the well, convert the higher molecular weight aromatic compounds into lower molecular weight compounds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows, by means of a block diagram, the different stages the biotechnological process of the present invention comprises:

-   -   a) Sampling and characterization,     -   b) Formulation of the culture media,     -   c) Obtaining of indigenous microorganisms,     -   d) Production of the IMP culture,     -   e) Production of the inocula for oil recovery, and     -   f) Oil recovery tests in porous media.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a biotechnological process to enhance recovery of oil with an API Gravity from 14 to 25, contained in carbonate-containing rock and/or clayey sandstone porous systems, presenting a permeability from 7 to 100 mD. The process of the invention is suitable for oil reservoirs associated to zones with a low oil recovery factor. The process of the invention uses the extremophile microorganisms activity: thermophile, barophile, halotolerant, acidotolerant and anaerobes that are indigenous from the oil reservoirs as well as its metabolites, such as surfactants, gases, acids and solvents, thereby improving the mobility of oils.

The biotechnological process of the present invention is based on the study of the rock characteristics and fluids from the reservoir, the formulation of culture media based on the reservoir study, obtaining and characterization of indigenous microorganisms from the petroleum reservoir, as well as the kinetic evaluation of metabolite production useful in the oil recovery.

The microorganisms and the substrates are incorporated into the well. After an incubation period from 5 to 10 days, the microorganisms and the synthesized metabolites increase the oil recovery. These anaerobe microorganisms can develop at temperatures ranging from 60 to 95° C., pressures from 7 to 154.6 Kg/cm² (100 to 2,200 psi) and NaCl content from 5,000 to 45,000 ppm.

To help understand better the biotechnological process of the present invention FIG. 1 shows, by means of a block diagram, the different stages comprising:

-   a) Sampling and characterization.—This step includes collecting     samples from the rock and fluids from the reservoir (oil and water),     for their characterization and determining oil API Gravity, type of     porous media (clayey sandstone and/or carbonate-containing rock),     rock porosity and permeability, as well as the microbial growth     conditions: such as temperature, pressure and salinity. The samples     from the reservoir must be kept at 4° C. for preservation until     performing the analysis; -   b) Formulation of the culture media.—This step includes determining     the formulation of nutrients, for the extremophile microorganisms     used in the present invention, where the formulation comprises the     components from Table 1 and the formulation of trace elements from     Table 2:

TABLE 1 Nutrients formulation from the culture media Compound Concentration, (g/l) NaCl  5.0-45.0 NH₄Cl  0.1-0.50 K₂HPO₄  0.1-0.40 MgCl₂ 6H₂O 0.5-1.5 CaCl₂ 2H₂O 0.05-0.2  KCl 0.2-0.4 NiCl₂ 6H₂O 0.025-0.05  Yeast extract 1.0-4.0 Tryptone peptone 1.0-4.0 Cisteine-HCl 0.1-0.5 Molasses  2.0-15.0 Resazurin 0.1% 1.0 (ml/l) Trace elements    5.0-10.0 (ml/l)

TABLE 2 Trace elements formulation like nutrients from the culture media Compound Concentration, (g/l) MgSO₄ 2.0-3-0 MnSO₄ 0.5-1.0 NaCl 1.0-2.0 FeSO₄ 0.1-0.2 CoSO₄ 0.1-0.2 CaCl₂ 0.1-0.2 ZnSO₄ 0.1-0.2 CuSO₄ 0.01-0.05 B(OH)₃ 0.01-0.03 Na₂MoO₄ 2H₂0 0.01-0.03 NiCl₂ 0.025-0.05 

This stage also comprises the mineral media formulation, its sterilizing conditions are 1.05 Kg/cm² (15 psi) and 122° C. for 22 minutes, the order of additions of components, its maintenance in anoxic conditions and the adjustment of the hydrogen potential;

-   c) Obtaining indigenous microorganisms.—This step includes the     selection, activation and preservation of extremophile     microorganisms obtained from the 14 to 25° API oils, capable of     producing useful metabolites to enhance oil recovery: This stage     includes:     -   Obtaining microorganisms from oil samples, cultured in a media         formulated for the development of useful organisms for oil         recovery, obtaining anaerobe cultures adapted to temperature         conditions from 60 to 95° C., salinity from 5,000 to 45,000 ppm         a NaCl content, and pressure from 7 to 154.6 Kg/cm² (100 to         2,200 psi) from the reservoir and/or well, and which can develop         in the presence of oils from 14 to 25° API;     -   From the preserved samples of the collected fluids         microorganisms are obtained. The microbiological processes are         carried out in anaerobic conditions, with N₂ atmosphere in all         the steps, temperatures from 60 to 95° C., preferably from 70 to         80° C., and salinities from 5,000 to 45,000 ppm and NaCl         content, in mineral media, complying with the nutritional         requirements of the microorganisms. Different carbon sources are         evaluated such as: molasses, glucose and sucrose at different         concentrations;     -   The different microbial groups present in the system are         determined such as fermentatives, methanogen, sulphate reducers,         among others. The balance of nutrients is determined (media         formulation) and the most suitable salinity conditions for the         production of metabolites useful to enhance oil recovery is         determined. The selection of cultures is carried out based on         its activity to produce surfactants, gases, acids and solvents,         among others;     -   Cultures are stabilized by means of periodical reseeding at         temperatures from 60 to 95° C. and salinity from 5,000 to 45,000         ppm;     -   The preservation of the microorganisms, in which cultures are         preserved by means of microorganisms fixation to an inert         support, in atmosphere of N₂ and in sterile conditions. The         effectiveness of the preservation method is determined by         reseeding in fresh media fixed microorganisms. -   d) Production IMP culture.—This step includes preparing the culture     media for stimulating microorganisms useful to enhance oil recovery     from oil samples, adapted to temperature conditions from 60 to 95°     C., preferably from 70 to 80° C., salinity from 5,000 to 45,000 ppm     with NaCl content and pressure from 7 to 154 Kg/cm² (100 to 2,200     psi) from the reservoir and/or well, able to develop in the presence     of oils from 14 to 25° API:     -   The microbial culture made up by micro organisms: thermophiles,         barophiles, halotolerants, acidotolerants, and anaerobes;         containing the following micro organisms: Thermoanaerobacter         ethanolicus, Thermoanaerobacter uzonensis, Thermoanaerobacter         inferii, Geothermobacterium sp, Methanobacterium subterraneum,         Methanobacterium formicicum, Methanolinea tarda and/or         Methanoculleus sp; -   e) Inoculum production for oil recovery.—The inoculum is made up by     indigenous microorganisms from the reservoir: stage c), and the IMP     culture from the stage d); -   f) Oil recovery tests in porous media:     -   Non consolidated Systems.—This step includes the addition of the         microbial culture (IMP culture and indigenous microorganisms)         and nutrients to the porous media oil saturated and recovery         evaluation, (Example 1).     -   Consolidated Systems (core).—This step includes the addition of         the microorganisms solution (IMP culture and indigenous         microorganisms) and the media formulated to the porous media,         along with the water injection, for a confinement period from 5         to 10 days, leading to the microbial development and the         metabolites synthesis in the system to promote oil mobility and         recovery (Examples 2 to 5); and     -   Well.—This step includes the biotechnological process in the         well, to enhance hydrocarbon recovery, additional to the         secondary recovery process, (Example 6).

The biotechnological process of the present invention comprises biostimulation and bioaugmentation of microorganisms in low permeability porous media, from 7 to 100 mD, at temperatures from 60 to 95° C., preferably from 70 to 80° C., pressures from 7 to 154.6 Kg/cm² (100 to 2,200 psi), salinity from 5,000 to 45,000 ppm and NaCl content, in anaerobic conditions.

The metabolites obtained during the biotechnological process in the present invention includes surfactants, gases (carbon dioxide, methane and hydrogen) acids (acetic, butyric and propionic), solvents (ethanol and acetone) to enhance oil mobility from 14 to 25° API, through mechanisms such as pressurization, dissolution, emulsification and viscosity reduction.

The biotechnological process of the present invention can be applied to porous media, clayey sandstone and/or carbonate-containing rock with 15 to 25% porosity.

The biotechnological process of the present invention leads to an oil recovery up to 30%, additional to the secondary recovery process in porous media.

EXAMPLES

Some practical examples of the present invention are following stated for a better understanding, not limiting its scope.

Example 1 Oil Enhanced Recovery In Non Consolidated Porous Media Using Microorganisms

Oil recovery was evaluated, using porous media obtained from clayey sandstone and/or carbonate-containing rocks. The rocks were ground, meshed and selected, based on the corresponding size, from 200 to 500 μm. 30 g of rock were added per system and covered, sealed and sterilized. The rocks were impregnated up to saturation with oils from 14 to 25° API using vacuum in a N₂ atmosphere. Under sterile conditions, the mineral media (Tables 1 and 2), the carbon source, indigenous microorganisms from oils and the IMP culture were added. The systems were incubated at 60-95° C. during 5 to 10 days. The gas production, surfactants, solvents and acids, sugar consumption and oil recovery were determined.

In the tests performed microorganisms favored oil release, obtaining a recovery above 24%.

The presence of metabolites was detected in all inoculated systems, (surfactants, gases, acids and solvents), promoting the mobility of oils from 14 to 25° API.

Example 2 Oil Recovery Tests in Consolidated Porous Media Using Microorganisms

For oil recovery experimental tests, carbonate-containing nucleus were used (10×5 cm), as well as oil from Mexican fields (14-25° API). Core fragments used (10×5 cm) in the experimental cell were rinsed with solvents and vacuum dried at 80° C. temperature. The core composition is synthesized in Table 3.

TABLE 3 Chemical composition or carbonated rocks Compound Rock composition % CaCO₃ 97.3 MgCO₃ 0.45 Al₂O₃ 0.55 SiO₂ 1.7

The experimental system consisted of a continuous injection pump, an oven for temperature control, transference cylinders, a differential pressure transducer, a data acquisition system and an experimental cell.

The experimental cell containing the core, was placed inside an oven for temperature control from 60 to 95° C. Transference cylinders were used to displace fluids (brine, oil and inocula) using and injection pump. A differential transducer was installed to monitor pressure drop. A system was finally set up for gas sampling in the outlet of the experimental cell.

The core fragment was placed inside a rubber sleeve in the experimental cell. Both were installed in the inlet and outlet diffusers. Confinement pressure was held between 7 and 154.6 Kg/cm² (100-2,200 psi) during the experiment. The experimental cell was installed inside the displacement system, which was held at a vacuum for an hour. The brine along with the injection flow from 5 to 10 ml/h was later displaced, up to a constant differential pressure; once the core was saturated, its porosity and permeability were determined, using different brine inlet flows. The core fragment was later saturated with oil from 14 to 25° API, at a 5 to 10 ml/h flow to reach an approximate oil saturation of 80%. Secondary oil recovery was carried out by means of brine injection at a 5 to 10 ml/h flow.

Once the secondary recovery process was concluded, the mineral media was injected with the carbon source, the inocula and the culture media to a 5 to 10 ml/h flow, until reaching 1.5-3.0 porous volume. The system was held in confinement from 60 to 95° C. from 5 to 10 days: during this period microorganisms grew producing metabolites (acids, surfactants, solvents and gases), useful to enhance oil mobility and recovery. The last phase of enhanced recovery by means of microbial, consisted on brine injection to a 5 to 10 ml/h flow, thus displacing oil released by microorganisms effect. The oil was recovered and quantified at the system outlet.

An oil recovery from 18.17 to 20.2% by volume of the original volume of core saturation from the microbial activity, additional to that through the secondary recovery process.

Experimental conditions are reported in Table 4.

TABLE 4 Oil recovery conditions from Examples 2 and 4 Characteristics Example 2 Example 4 Microbian culture IMP¹ and IMP¹ and Indigenous Indigenous API Gravity 14 to 25 14 to 25  Kinematic viscosity 30 to 50 30 to 50  30° C., mPa s⁻¹ Brine, % NaCl  5 to 50 5 to 50 Type of rock Carbonate- Clayey sandstone containing Length, cm 10   14   Diameter, cm 10   5  Temperature, ° C. 60 to 95 60 to 95  Presure, Kg/cm² 7 to 154.6 7 to 154.6 (psi) (100 to 2,200) (100 to 2,200) Porous volume, cm³ 35.7  208   Porosity, % 18.1   19.73 Permeability, mD  7 to 15 90 to 100 Flow, ml/h  5 to 10 5 to 10 initial oil saturation, 78.43 70.2 % Brine saturation, % 21.57 19.8 Brine injection recovery 30.47 to 33.53 45.3 to 49.45 (secondary), % Biotechnological process 18.17 to 20.23 20.6 to 25.4  recovery of the present invention², % Total recovery % 48.6 to 53.7 65.9 to 74.89 (secondary plus biotechnological process) Detected metabolites Acids, gases, Acids, gases, surfactants and surfactants and solvents solvents ¹The indigenous culture is made up of microorganisms isolated from oil from a Mexican petroleum well. ²Recovery was due to a biotechnological process application of the present invention, additional to that obtained in the secondary recovery process (water injection).

Example 3

All of the conditions used were identical to those from Example 2, except for the microorganisms and nutrients addition, which was carried out with 3 injection cycles and short confinement periods from 5 to 10 days. The oil recovery obtained by means of the biotechnological process from the present invention was from 20.1 to 24.3%, additional to that obtained through the secondary recovery process.

Example 4

All conditions used were identical to those from Example 2, except for the use of Berea sandstone, (14×10 cm), having a chemical composition is described in Table 5:

TABLE 5 Chemical composition of the Berea sandstone rock Composition of the Compounds sandstone rock (%) SiO₂ 93.13 Al₂O 3.86 Fe₂O₃ 0.11 FeO 0.54 MgO 0.25 CaO 0.10 Others 2.01

An oil recovery from 20.6 to 25.4% due to microbial activity, was obtained from the initial volume of the core saturation, additional to that obtained through the secondary recovery.

Example 5

All the conditions used were identical to those from Example 3, except for the nutrients and microorganisms addition, which was carried out with 3 injection cycles and confinement short periods from 5 to 10 days, obtaining an oil recovery by the biotechnological process of the present invention from 25.1 to 29.9%, additional to that obtained by the secondary recovery process.

Example 6

To carry out the oil recovery test in a well, a culture media was prepared, using the following nutrients, at the conditions stated in g/l: NaCl (15-35), NH₄Cl (0.2-0.4), K₂HPO₄ (0.1-0.3), MgCl₂ 6H₂O (0.7-1.3), CaCl₂ 2H₂O (0.05-0.1), KCl (0.25-0.3), NiCl₂ 6H₂O (0.03-0.04) yeast extract (1.0-3.0), triptone peptone (1.0-3.0), cisteine-HCl (0.1-0.3), molasses (4.0-10.0), resazurin 0.1% (1.0 ml/l), and trace elements solution (6.0-8.0 ml/l, according to Table 2).

The culture media was sterilized with heat at 122° C., 1.05 Kg/cm² (15 psi), for 22 minutes. The culture media was held in anaerobic conditions in suitable cells for the required volume, according to the test area and associated facilities. Indigenous cultures from the reservoirs and the IMP culture were added, selected by its capacity to produce metabolites like acids, surfactants, solvents and gases, enhancing the oil recovery conditions.

The microorganisms and nutrients were prepared ex situ. They were confined in suitable reservoirs to guarantee its stability during transportation to the application point. Transfer cylinders were used to move the fluids (nutrients and inocula) using an injection pump. The monitoring system included a differential transducer to monitor pressure drop and a sampling system was implemented.

The culture media, the inoculum, the carbon source and the water were introduced through pumping into the well. The well was closed from 5 to 10 days. The microorganisms produced the metabolites carbon dioxide, methane, hydrogen, surfactants, ethanol, acetic acid, thereby favoring oil recovery.

The oil increase obtained in the recovery by the microbial process was 11.3%, additional to that obtained by the secondary recovery process.

Example 7

For a better comprehension of the resulting effect from the microorganisms activity used in the biotechnological process of the present invention, regarding the composition of hydrocarbon fractions used in Example 3, in the following hydrocarbon fractions are shown in Table 6:

-   1. Initial oil.—oil from the petroleum well such as the one used in     Example 3; -   2. Impregnation oil.—Effluent resulting from oil saturation in the     core, as stated in Example 3; -   3. Secondary recovered oil.—Effluent obtained as of the recovery     process by means of brine injection, as stated in Example 3; and -   4. Microbial recovered oil.—Oil from microbial recovery process, as     stated in Example 3.

TABLE 6 SARA Analysis of oil used in Example 3 Oil Sample Saturates Resins Aromatics Asphaltenes Initial oil 19.81 12.57 58.9 8.72 Impregnation oil 19.55 18.63 54.27 7.55 Secondary recovered oil 18.59 18.85 54.92 7.64 Microbial recovered oil 29.81 18.57 42.86 8.76

Results from Table 6 show that with the biotechnological process of the present invention, microorganisms were able to modify the aromatic hydrocarbons turning them into aliphatic (saturates) or aromatic compounds with a lower molecular weight:

-   -   Aromatic compounds in the all types of oils except the initial         oil, decreased after incubation and recovery, decrease in the         “microbial recovered oil” was more important in a 16%, implying         that the microorganisms present in the process of this Example         have the capacity to degrade this type of compounds.     -   Regarding the saturated fraction, there were no significant         differences in the initial oil with “Impregnation oil” and         secondary recovered oil, however in the “microbial recovered         oil” oil there was a 10% increase; being related to the         aromatics degradation. 

1. A biotechnological process for oil recovery in low permeability porous media, comprising the following stages: a) Sampling and characterization of samples collected from rock and fluids from the reservoir, for their characterization, and measuring the temperature, pressure and salinity conditions in the reservoir; b) Producing a culture media formulation containing a nutrient formulation for microorganisms development from the samples in stage a), c) Obtaining indigenous microorganisms including extremophile microorganisms selection, activation and preservation, from 14 to 25° API oils, that are able to produce metabolites useful for oil recovery, and can develop at 60 to 95° C., temperatures and NaCl content from 5,000 to 45,000 ppm, pressures from 7 to 154.6 Kg/cm² (100 to 2,200 psi) in anaerobic conditions, with mineral media Table 2 and 3), d) Producing an IMP culture including a microbial culture adapted to 60 a 95° C. temperature conditions, 5,000 a 45,000 ppm salinity of NaCl content, and 7 to 154.6 Kg/cm² pressure (100 to 2,200 psi) from the reservoir and/or well developing in the presence of 14 to 25° API oils and containing at least one of the microorganisms selected from the group consisting of Thermoanaerobacter ethanolicus, Thermoanaerobacter uzonensis, Thermoanaerobacter inferii, Geothermobacterium sp, Methanobacterium subterraneum, Methanobacterium formicicum, Methanolinea tarda and Methanoculleus sp; e) Producing an inoculum for oil recovery where the inoculum is made up of said indigenous microorganisms from the oil reservoir stage c), and the IMP culture of stage d), f) Oil recovery tests in porous media comprising the addition of the culture media of stage b) and the inoculum of stage e) to the porous media, along with the water injection; confinement from 5 to 10 days for metabolites production favoring oil mobility and recovery; and increase oil recovery, additional to the secondary recovery process.
 2. A biotechnological process of claim 1, wherein said oil is located in porous media with the following conditions; from 7 to 100 mD permeability, 60 to 95° C. temperature, 7 to 154.6 Kg/cm² (100 to 2,200 psi) pressure, 5,000 to 45,000 ppm salinity (NaCl).
 3. A biotechnological process of claim 1, wherein porous media are preferably clayey sandstone and or carbonate-containing rock.
 4. A biotechnological process of claim 1, wherein the porous media has a 15 to 25% porosity.
 5. A biotechnological process of claim 1, wherein the indigenous extremophiles microorganisms from oil reservoir are thermophile, barophile, halotolerant and acidotolerant, and able to grow at 60 to 95° C. temperatures, pressures from 7 to 154.6 Kg/cm² (100 to 2,200 psi) and NaCl content from 5,000 a 45,000 ppm, in anaerobic conditions.
 6. A biotechnological process of claim 1, wherein nutrient formulation for extremophile microorganisms, stated in stage b), comprises: Nutrient formulation from the culture media Compounds Concentration (g/l) NaCl  5.0-45.0 NH₄Cl  0.1-0.50 K₂HPO₄  0.1-0.40 MgCl₂ 6H₂O 0.5-1.5 CaCl₂ 2H₂O 0.05-0.2  KCl 0.2-0.4 NiCl₂ 6H₂O 0.025-0.05  Yeast extract 1.0-4.0 Triptone peptone 1.0-4.0 Cisteine HCl 0.1-0.5 Molasses  2.0-15.0 Resazurin 0.1% 1.0 (ml/l) Trace elements 5.0-10.0 (ml/l)

Trace elements formulation as nutrient of the culture media Compounds Concentration (g/l) MgSO₄ 2.0-3-0 MnSO₄ 0.5-1.0 NaCl 1.0-2.0 FeSO₄ 0.1-0.2 CoSO₄ 0.1-0.2 CaCl₂ 0.1-0.2 ZnSO₄ 0.1-0.2 CuSO₄ 0.01-0.05 B(OH)₃ 0.01-0.03 Na₂MoO₄ 2H₂0 0.01-0.03 NiCl₂ 0.025-0.05 


7. A biotechnological process of claim 1, wherein the indigenous culture, stated in stage c), is made up by microorganisms producing metabolites: surfactants, gases, acids and solvents, to improve oil mobility of oil having 14 to 25° API.
 8. A biotechnological process of claim 1, wherein the carbon source, stated in stage c), is molasses, glucose or sucrose.
 9. A biotechnological process of claim 1, wherein the temperature in which extremophile microorganisms are developed, in stages c) and d), is from 70 to 80° C.
 10. A biotechnological process of claim 1, wherein cultures in stage c) are preserved by means of fixed microorganisms in an inert support, N₂ atmosphere and in sterility conditions.
 11. A biotechnological process of claim 1, wherein the addition of the culture media to the porous media or well, along with a water injection, stated in stage f), is performed in a confinement period from 5 to 10 days, in one or in several cycles leading to the microbial development and the metabolites synthesis in the system.
 12. A biotechnological process of claim 1, wherein up to 30% of oil is recovered in addition to the secondary recovery process in porous media. 